Antibodies against n-procalcitonin

Inactive Publication Date: 2013-02-21
UNIV DE SEVILLA +1
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  • Abstract
  • Description
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Benefits of technology

[0091]On the other hand, the role of endogenous N-PCT was studied in LPS-induced mortality. N-PCT was specifically neutralized with an anti-N-PCT antibody 1 hour before LPS administration. Survival was analysed during 7 days. Death occurred from 8 to 48 hours after LPS administration. As shown in FIG. 6, the passive immunization of rats with a single dose of anti-M-PCT increased the survival rate 100% in comparison with 80% in animals treated with IgG control, from 8 to 12 hours after LPS administration. After 18 hours, the survival was 60% in control rats compared with 85% of rats treated with anti-N-PCT. After 2 days the death in the control rats was 100%, whilst in the rats treated with anti-N-PCT the death was 15% from the first day until the end of the study.
[0092]The rats that survived 2 days showed an improvement from that time on, represented by greater mobility and they did not shown lethargy or diarrhoea. 7 days after administration. None of the animals treated with anti-N-PCT showed signs of systemic infection. This suggests that N-PCT has a relevant role in the response that led to survival in the event of septic shock.
[0093]Later, experiments were developed to determine if the circulating N-PCT regulates the balance between the pro and anti-inflammatory cytokines. In animals treated with anti-N-PCT the levels of cytokines TNF-α, IL-1β, IL-10, as well as of PCT returned to basal situation in comparison with the untreated rats. These changes are associa

Problems solved by technology

Despite this, the physiological role of PCT as well as its systemic effects are not well known.
These results do not justify its supposed role as secondary mediator in sepsis.
This detection system is very non-specific and may also require the use of two antibodies.
It also has the disadvan

Method used

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Examples

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Effect test

example 1

Effects of the Intracerebroventricular Administration of Anti-N-PCT in N-PCT-Induced Anorexia

[0084]The effect of the cerebroventricular administration in the intake of is shown in FIG. 1. The intake was measured 2, 4 and 24 hours after a single intracerebroventricular injection of N-PCT (5 μg), at the start of night-time feeding. In accordance with previous data, the icy administration of N-PCT reduced food intake during 24 hours. In comparison with the administration of the carrier, N-PCT reduced the intake by 78% during the first 2 hours and 83% during the first 4 hours (P<0.001). The accumulated intake during 24 hours was reduced 26% with respect to the controls. The treatment with anti-NPCT reversed these effects (p<0.001). This confirms the specificity of the antibodies used. Furthermore, these data demonstrate the implication of NPCT in metabolic stress.

example 2

Effects of a Lethal Dose of LPS on the Behaviour and Plasma Levels of ACTH, CT and Cytokines

[0085]After the administration of a high dose of LPS (15 mg / kg, ip), all rats showed serious symptoms of disease. They quickly became lethargic and anorexic and showed standing on end of the hair, chromodacryorrhoea and diarrhoea. Many of the animals showed a short hypothermia followed by fever. The animals seemed moribund after 12 hours and none survived 48 hours.

[0086]As shown in FIG. 2A, the administration of 15 mg / kg of LPS caused a decrease in body temperature (Tb) from 30 minutes after the administration of LPS, and reached a maximum after one and a half hours. (36.5±0.06° C.). Within the first 2 hours after LPS administration, Tb began to gradually increase, reaching a maximum after 8 hours (38.67±0.15° C.), which lasted 12 hours (37.49±0.12° C.). Tb was not altered by the administration of PFS. The plasma levels of ACTH, CT, TNF-α, IL-1β and IL10 were also measured after the injection...

example 3

Expression of the Calca-1 Gene and Production of PCT in Plasma and Hypothalamus After LPS Administration

[0087]The Calca-1 gene was expressed at high levels in the hypothalamus after LPS administration (FIG. 3A). Similarly to PCT levels, the expression of Calca-1 began its increase one hour after administration. It reaches the maximum after 3 hours and begins to decrease after 6 hours, reaching basal levels after 12 hours. There is a low expression of Calca-1 in the rats treated with PFS.

[0088]LPS administration produces a fast and striking increase in PCT levels in plasma and hypothalamus. Between 1 and 3 hours. They reach a maximum after 12 hours and are maintained until the death of the animal. The increases are of 6000 and 540 times after 12 hours in plasma or hypothalamus, respectively. Those increases of PCT in plasma correlate with the production of PCT in the hypothalamus. There appears a fast increase after one hour that is maintained over time.

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Abstract

Specific antibodies against N-procalcitonin, peptides, genetic constructions and methods for the obtainment of the peptides used in the obtainment of the antibodies. These antibodies can be used for preparing drugs, or diagnostic kits for diseases that develop with systemic inflammatory response or metabolic stress.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 61 / 499,808, filed Jun. 22, 2012 and entitled “Antibodies Against N-Procalcitonin” in the name of Eva TAVARES VÁZQUEZ et al., which is incorporated by reference herein in its entirety.FIELD[0002]The present invention is within the field of biotechnology and medicine. It relates to antibodies against N-procalcitonin, peptides, genetic constructions and the methods for the obtainment of the peptides used in the obtainment of the antibodies, and the uses of said antibodies.PRIOR ART[0003]Procalcitonin (PCT or proCT) is a glycopeptide hormone of 116 amino acids and approximately 13 kDa of molecular weight. This molecule is the precursor of calcitonin. Its synthesis starts with the transcription of the Calca-1 gene situated in chromosome 11p. This subscript is subsequently processed giving rise to preprocalcitonin, precursor of PCT. This precurso...

Claims

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Application Information

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IPC IPC(8): C07H21/04
CPCC07K14/585A61K2039/505C07K16/26
Inventor TAVARES VÁZQUEZ, EVAMIÑANO SÁNCHEZ, JAVIER
Owner UNIV DE SEVILLA
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